|
| Status |
Public on May 18, 2007 |
| Title |
Thymus, mouse17K-B, 04 AMC33 F 193 C |
| Sample type |
RNA |
| |
|
| Source name |
Mus musculus thymus
|
| Organism |
Mus musculus |
| Characteristics |
Strain: C57BL/6, Tissue: thymus
Gender: female, Age: 193 days Diet: Caloric restriction
|
| Treatment protocol |
100% regular feed to 13 weeks of age, 90% fortified feed for 1 week, 75% fortified feed for 1 week, then 60% fortified feed until the age of sacrifice
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Qiagen RNeasy Mini Kit protocol for animal tissues after disruption using a mini-bead beater-8 and 1.0mm glass beads.
|
| Label |
33P-dCTP
|
| Label protocol |
~5 ug of total RNA was radiolabeled with 33P-dCTP using Superscript II from Invitrogen. Labeled cDNA was separated from unincorporated probe and small nucleotides using a Bio-Rad size exclusion column, Bio-Spin 30 with SSC. 1ul of each sample was counted in a scintillation counter and the remainder was used for hybridization.
|
| |
|
| Hybridization protocol |
Radiolabeled cDNA was denatured at 95 degrees C for 7 minutes and added to 12 mls of Invitrogen microhyb containing 10% dextran sulfate with Cot-1 DNA (Invitrogen) and Poly-A DNA oligos, (Sigma) added as blocking agents. Arrays were hybridized overnight at 55 degrees C to mouse 17K-B cDNA filters. The hybridized filters were quickly rinsed once and then washed twice at 65 degrees placed onto steel plates covered with plastic wrap and exposed to Kodak storage Phosphorimaging screens for 3-4 days.
|
| Scan protocol |
Imaging screens were scanned at 50um resolution in a Storm Phosphorimager.
|
| Description |
04 AMC33 F 193 C
|
| Data processing |
All data was extracted using ArrayPro Software and processed in Excel spreadsheets using Z normalization of each array. Differently expressed genes were identified between the different sample groups by z-ratio analysis using an in-house microarray analysis program.
|
| |
|
| Submission date |
May 17, 2007 |
| Last update date |
Jun 22, 2020 |
| Contact name |
Supriyo De |
| Organization name |
NIA-IRP, NIH
|
| Department |
Laboratory of Genetics and Genomics
|
| Lab |
Computational Biology & Genomics Core
|
| Street address |
251 Bayview Blvd
|
| City |
Baltimore |
| State/province |
Maryland |
| ZIP/Postal code |
21224 |
| Country |
USA |
| |
|
| Platform ID |
GPL782 |
| Series (1) |
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